Department of Radiation Oncology, University of Michigan, Ann Arbor, MI 48109, USA.
Anal Biochem. 2011 May 1;412(1):1-8. doi: 10.1016/j.ab.2011.01.028. Epub 2011 Jan 27.
The receptor tyrosine kinase c-Met and its ligand, hepatocyte growth factor/scatter factor (HGF/SF), modulate signaling cascades implicated in cellular proliferation, survival, migration, invasion, and angiogenesis. Therefore, dysregulation of HGF/c-Met signaling can compromise the cellular capacity to moderate these activities and can lead to tumorigenesis, metastasis, and therapeutic resistance in various human malignancies. To facilitate studies investigating HGF/c-Met receptor coupling or c-Met signaling events in real time and in living cells and animals, here we describe a genetically engineered reporter where bioluminescence can be used as a surrogate for c-Met tyrosine kinase activity. c-Met kinase activity in cultured cells and tumor xenografts was monitored quantitatively and dynamically in response to the activation or inhibition of the HGF/c-Met signaling pathway. Treatment of tumor-bearing animals with a c-Met inhibitor and the HGF neutralizing antibody stimulated the reporter's bioluminescence activity in a dose-dependent manner and led to a regression of U-87 MG tumor xenografts. Results obtained from these studies provide unique insights into the pharmacokinetics and pharmacodynamics of agents that modulate c-Met activity and validate c-Met as a target for human glioblastoma therapy.
受体酪氨酸激酶 c-Met 及其配体肝细胞生长因子/分散因子（HGF/SF）调节细胞增殖、存活、迁移、侵袭和血管生成所涉及的信号级联反应。因此，HGF/c-Met 信号的失调会损害细胞调节这些活动的能力，并可能导致各种人类恶性肿瘤的肿瘤发生、转移和治疗耐药性。为了促进研究实时和在活细胞和动物中研究 HGF/c-Met 受体偶联或 c-Met 信号事件，我们在这里描述了一种遗传工程报告基因，其中生物发光可以作为 c-Met 酪氨酸激酶活性的替代物。通过监测培养细胞和肿瘤异种移植物中 c-Met 激酶活性，对 HGF/c-Met 信号通路的激活或抑制进行定量和动态监测。用 c-Met 抑制剂和 HGF 中和抗体处理荷瘤动物以剂量依赖性方式刺激报告基因的生物发光活性，并导致 U-87 MG 肿瘤异种移植物的消退。这些研究的结果提供了对调节 c-Met 活性的药物的药代动力学和药效动力学的独特见解，并验证了 c-Met 作为人类胶质母细胞瘤治疗靶点的适用性。
